LOCUS       sequence-120554-        3236 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MoClo Basic Part: CDS - Controller protein, lacI repressor (in 
            concert with CAP, represses pLacI, R0010). Modified to fix illegal 
            site. [C:C0012m:D].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3236)
  AUTHORS   Iverson SV, Haddock TL, Beal J, Densmore DM
  TITLE     CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part 
            Library Enable Rapid Combinatorial Design for Synthetic and 
            Traditional Biology.
  JOURNAL   ACS Synth Biol. 2016 Jan 15;5(1):99-103. doi: 
            10.1021/acssynbio.5b00124. Epub 2015 Nov 4.
  PUBMED    26479688
REFERENCE   2  (bases 1 to 3236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.5b00124"; journalName: "ACS Synth Biol"; date: 
            "2016-01-15- 15"; volume: "5"; issue: "1"; pages: "99-103"
COMMENT     Sequence Label: C0012m (lacI)_CD
FEATURES             Location/Qualifiers
     source          1..3236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             14..1093
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(33..52)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1094..1126
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     misc_feature    1144..1164
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1170..1187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1341..1929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1421..1440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2100..2891,2892..2960))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2723..2742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2961..3065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3133..3151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3155..3198)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3201..3222
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aatgatggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
       61 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
      121 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
      181 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
      241 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
      301 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
      361 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
      421 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
      481 caacagtatt attttctccc atgaggacgg tacgcgactg ggcgtggagc atctggtcgc
      541 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
      601 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
      661 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
      721 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
      781 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
      841 cgaagatagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
      901 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
      961 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     1021 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1081 ggaaagcggg caggctgcaa acgacgaaaa ctacgcttta gtagcttaat aaaggtagag
     1141 acctactagt agcggccgct gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1201 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1261 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1321 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat
     1381 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1441 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1501 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1561 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1621 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1681 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1741 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1801 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1861 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1921 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1981 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2041 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2101 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2161 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2221 agtgctgcaa tgataccgcg cgacccacgc tcaccggctc cagatttatc agcaataaac
     2281 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2341 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2401 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2461 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2521 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2581 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2641 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2701 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2761 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2821 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2881 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2941 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3001 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3061 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3121 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     3181 ttcgctaagg atgatttctg gaattcgcgg ccgcttctag agactagtgg gtctca
//
